DNA条形码技术在小花蝽属昆虫分类鉴定中的应用

本研究利用DNA条形码技术对13种小花蝽属Orius Wolff昆虫进行了鉴定,进行了59条COI基因序列碱基组成及种内、种间遗传距离的分析,采用邻接法、最大简约法、贝叶斯推论法构建了系统发育树。结果表明,小花蝽属昆虫COI基因序列碱基组成与典型的昆虫线粒体DNA一致,A+T平均含量(66.4%)明显高于G+C含量,密码子的第3位A+T含量高达90.7%,碱基替换多为同义替换;13种小花蝽种内平均遗传距离为0.008,种间平均遗传距离为0.128,种内、种间遗传距离没有重叠区域。3种方法构建的系统发育树的聚类分析与形态学鉴定结果基本一致,除微小花蝽Orius minutus(Linnaeus)可能存在隐存种现象外,其他同一种群的不同个体单独聚为一支。利用DNA条形码技术对小花蝽属昆虫进行物种快速分子鉴定具有可行性。     

基于线粒体COⅠ基因的DNA条形码技术在昆虫分子鉴定中的应用

本文对基于线粒体COⅠ基因的DNA条形码技术在昆虫分子鉴定中的应用进行了综述,对DNA条形码的意义、DNA条形码的原理、线粒体COⅠ基因的特点、DNA条形码的操作、DNA条形码技术在昆虫分子鉴定中的应用研究进展、DNA条形码的发展趋势以及有关DNA条形码的争论进行了介绍.     

果园螟蛾总科部分种类DNA条形码鉴定

为检验DNA条形码在鳞翅目螟蛾总科蛾类鉴定中的可行性,对采自山西省太原市晋源区螟蛾总科26种78头蛾类标本分别提取了DNA,扩增了全部78头标本的线粒体cox1基因和其中75头标本的核糖体28S基因,并通过构建系统发育树、计算遗传距离及种间差异阈值等方法,对所有标本进行了鉴定和比较分析,检验了国际DNA条形码数据库BOLD(the barcode of life data)系统的鉴定成功率。结果表明,基于cox1基因和28S基因的系统发育树鉴定成功率分别为100.00%和97.14%,BOLD系统的鉴定成功率达到了67.94%。基于最大简约法、邻接法和最大似然法构建的系统发育树,鉴定结果均相同。基于cox1基因的种内遗传距离全部小于1.00%,种内种间的遗传距离形成明显的3.00%阈值现象。研究表明,cox1及28S基因均适用于供试螟蛾总科种类的鉴定,核糖体28S基因可以作为DNA条形码鉴定的辅助基因;BOLD系统数据库仍有待充实,且标本鉴定工作相对滞后;不同聚类分析方法对结果影响很小,其中邻接法计算速度快,更适合DNA条形码大数据的分析。     

基于mtCOⅠ基因的三种稻飞虱DNA条形码构建及应用

为构建白背飞虱、褐飞虱、灰飞虱3种稻飞虱DNA条形码,准确鉴定各虫态稻飞虱种类,随机选取35个供试虫体提取全基因组DNA,PCR扩增其中线粒体细胞色素氧化酶Ⅰ（mtCOⅠ）基因,分析所获得的基因组成特点,计算P距离值,构建系统发育树,并对稻飞虱若虫、卵进行种类鉴定。PCR扩增获得了长度为706 bp的稻飞虱mtCOⅠ基因片段,碱基组成特点种内、种间不同;3种稻飞虱的种间P距离值为25.53%~34.21%,种内为0~0.51%,种间值明显高于种内;系统发育树显示同种个体聚集在同一分支;同种卵、若虫的mtCOⅠ基因与已获得的成虫基因序列同源性最高值为100%,P距离值为0。研究表明,3种稻飞虱的DNA条形码可基于mtCOⅠ基因构建,且可准确鉴定各虫态。     

珠海口岸全国首次截获重要害虫鳄梨织蛾

2018年1月珠海检验检疫局首次从旅客携带的厄瓜多尔鳄梨中截获我国进口墨西哥、秘鲁鳄梨议定书中关注的检疫性有害生物——鳄梨织蛾(Stenoma catenifer),本文对该虫的寄主、分布及形态特征进行了详细描述。文中应用DNA条形码技术扩增了该虫的线粒体COⅠ序列,通过GenBank数据库比对分析,邻接法构建的系统发育树及BOLD数据库查询比对。结果与GenBank数据库中2条及BOLD数据库中11条鳄梨织蛾序列97%以上一致;构建的系统发育树显示截获样品与鳄梨织蛾聚为一支。因此,综合形态和分子数据确定所截获的有害生物为鳄梨织蛾。     

茶园茶黄蓟马及其近似种的DNA条形码鉴定

基于 mtDNA COI 基因的 DNA 条形码技术是一种新的物种鉴定手段,本文尝试应用该技术对6个不同地理种群的茶黄蓟马与茶园中的其他4种常见蓟马进行了鉴别。测定各种类 mtDNA COI 基因序列并在 BOLD 系统进行比对,采用 MEGA5构建系统进化树,结果表明基于 mtDNACOI 基因的DNA 条形码技术可实现对茶园中茶黄蓟马的快速鉴定。     

欧洲纵坑切梢小蠹的检疫鉴定研究

云浮口岸从土耳其进境木质包装中截获多头活小蠹虫,本研究对该小蠹虫的形态特征进行描述,同时采用小蠹科通用引物CI-J-2183和TL2-N-3014进行PCR扩增该虫的COⅠ基因序列。结果获得758 bp的基因序列,通过与Gen Bank数据库比对分析显示,该序列与Gen Bank数据库上的3条欧洲纵坑切梢小蠹COⅠ序列相似性高达99%;采用邻接法(NJ)构建的系统发育树也显示该虫与欧洲纵坑切梢小蠹亲缘关系极近且在同一进化分支中。结合形态特征和DNA条形码序列分析,可最终判定所截获昆虫为欧洲纵坑切梢小蠹。     

大穗看麦娘与日本看麦娘的DNA条形码鉴定

大穗看麦娘是我国麦田新发生的恶性杂草,与日本看麦娘苗期形态相近,导致难以识别和有效监测。本研究利用4个DNA 条形码候选序列(rbcL、matK、trnH-psbA和ITS2)对13份大穗看麦娘和10份日本看麦娘叶片材料进行分子鉴定,采用Vector NTI分析扩增的DNA序列峰图质量并比对碱基差异,通过MEGA 6.0软件中的K2P模型计算样本种内和种间的双参数遗传距离,采用邻接法构建系统发育树。结果表明,4个DNA 条形码序列中仅matK扩增测序结果不理想。日本看麦娘在4种DNA条形码序列中不存在种内差异,大穗看麦娘在rbcL、matK和ITS2序列中无种内差异,仅在trnH-psbA序列中存在7个差异位点。大穗看麦娘和日本看麦娘种间各DNA条形码序列均有差异,trnH-psbA、rbcL、matK和ITS2序列存在的差异位点数分别为6、3、14和28。ITS2的种间平均遗传距离大于rbcL,且具有特异性,适宜用于大穗看麦娘和日本看麦娘的准确鉴定。     

matK序列作为DNA条形码在苍耳属中的应用

以matK序列作为DNA条形码,对从国外截获的7种苍耳属植物进行物种区分鉴定研究,采用DNeasyOPlantMiniKit试剂盒进行总DNA的提取,应用通用引物对其matK基因进行扩增,测序得到7种苍耳属植物的marK序列,利用MEGA5．1软件对这7种苍耳属植物的matK序列进行比对和分析并构建系统树,结果显示：matK序列能从基因位点层面对苍耳属植物进行区分鉴定。     

景天属多肉植物基于ITS条形码鉴定探析

在多个条形码基因研究的基础上,本研究以ITS序列为目的片段通过DNA条形码技术对口岸截获的景天属多肉植物进行快速鉴定。采用通用引物扩增景天属10种32个供试样本的ITS片段并双向测序拼接,利用MEGA7软件进行序列比对,分析种内种间K2P距离并作barcoding gap,构建系统发育树。结果表明供试材料ITS序列的种内变异明显小于种间差异,种内种间变异存在明显的间隔区,每种的所有样品在NJ树上严格聚成单系,种间区分明显。初步判定ITS具有作为景天属多肉植物DNA条形码的潜在可能。     

Dermestid beetles in ‘evolution Canyon’, lower Nahal Oren, Mt. Carmel, including new records for Israel

Fifteen species of dermestid beetles were recorded at ‘Evolution Canyon’ (EC), Lower Nahal Oren, Mt. Carmel, Israel. They represent ~35% of known Israeli dermestid species. The following three species were recorded for the first time in Israel:Trogoderma svriaca Dalla Torre, 1911;Ctesias svriaca Ganglbauer, 1904; andAnthrenus (s.str.) jordaniens Pic, 1934. Adults of 13 species were collected on the more solar radiated, warmer and climatically more fluctuating south-facing slope (SFS); ten species were collected on the opposite, north-facing slope (NFS), which was cooler and climatically more stable. The abundance of adult dermestid beetles was 1.9 times higher on the SFS than on the NFS (86 and 47, respectively). Species richness and abundance distribution at EC (three collecting stations on each slope and one at the valley bottom) were significantly negatively correlated with the plant cover that consisted of trees and bushes (Spearmanr _s ,P=0.007 and 0.039, respectively) and perennials (Spearmanr _s ,P=0.039 and 0.077, respectively), indicating that non-woody plants were preferred by adult dermestid beetles.     

Epidemiology of Toxigenic Fungi and their Associated Mycotoxins for Some Mediterranean Crops

Recent data on the epidemiology of the common mycotoxigenic species of Fusarium, Alternaria, Aspergillus and Penicillium in infected or colonized plants, and in stored or processed plant products from the Mediterranean area are reviewed. Emphasis is placed on the toxigenicity of the causal fungal species and the natural occurrence of well known mycotoxins (aflatoxins, ochratoxins, fumonisins, trichothecenes, zearalenone, patulin, Alternaria-toxins and moniliformin), as well as some more recently described compounds (fusaproliferin, beauvericin) whose toxigenic potential is not yet well understood. Several Fusarium species reported from throughout the Mediterranean area are responsible of the formation of mycotoxins in infected plants and in plant products, including: Fusarium graminearum, F. culmorum, F. cerealis, F. avenaceum, F. sporotrichioides and F. poae, which produce deoxynivalenol, nivalenol, fusarenone, zearalenone, moniliformin, and T-2 toxin derivatives in wheat and other small grains affected by head blight or scab, and in maize affected by red ear rot. Moreover, strains of F. verticillioides, F. proliferatum, and F. subglutinans, that form fumonisins, beauvericin, fusaproliferin, and moniliformin, are commonly associated with maize affected by ear rot. Fumonisins, were also associated with Fusarium crown and root rot of asparagus and Fusarium endosepsis of figs, caused primarily by F. proliferatum. Toxigenic A. alternata strains and associated tenuazonic acid and alternariols were commonly found in black mould of tomato, black rot of olive and citrus, black point of small cereals, and black mould of several vegetables. Toxigenic strains of A. carbonarius and ochratoxin A were often found associated with black rot of grapes, whereas toxigenic strains of A. flavus and/or P. verrucosum, forming aflatoxins and ochratoxin A, respectively, were found in moulded plant products from small cereals, peanuts, figs, pea, oilseed rape, sunflower seeds, sesame seeds, pistachios, and almonds. Finally, toxigenic strains of P. expansum and patulin were frequently found in apple, pear and other fresh fruits affected by blue mould rot, as well as in derived juices and jams.     

A survey of the genera of<Emphasis Type="Italic">Microgaster</Emphasis> latreille and<Emphasis Type="Italic">Hygroplitis</Emphasis> thomson (Hymenoptera: Braconidae: Microgastrinae) from China

The genera ofMicrogaster Latreille 1804 andHygroplitis Thomson 1895 from China are presented systematically in this paper. Thirty-two species ofMicrogaster and three species ofHygroplitis are known in China. Diagnosis, character variation, distribution and host of each species among the two genera are presented, including its host and distribution. Keys to the species ofMicrogaster andHygroplitis are given. http://www.phytoparasitica.org posting Dec. 19, 2006.     

Plant Viruses Transmitted by Whiteflies

One-hundred and fourteen virus species are transmitted by whiteflies (family Aleyrodidae). Bemisia tabaci transmits 111 of these species while Trialeurodes vaporariorum and T. abutilonia transmit three species each. B. tabaci and T. vaporariorum are present in the European–Mediterranean region, though the former is restricted in its distribution. Of the whitefly-transmitted virus species, 90% belong to the Begomovirus genus, 6% to the Crinivirus genus and the remaining 4% are in the Closterovirus, Ipomovirus or Carlavirus genera. Other named, whitefly-transmitted viruses that have not yet been ranked as species are also documented. The names, abbreviations and synonyms of the whitefly-transmitted viruses are presented in tabulated form together with details of their whitefly vectors, natural hosts and distribution. Entries are also annotated with references. Whitefly-transmitted viruses affecting plants in the European–Mediterranean region have been highlighted in the text.     

Broad bean mottle virus in Morocco; curculionid vectors,and natural occurrence in food legumes other than faba bean (Vicia faba)

Broad bean mottle virus (BBMV) was transmitted from infected to healthy faba-bean plants by the curculionid weevilsApion radiolus Kirby,Hypera variabilis Herbst,Pachytychius strumarius Gyll,Smicronyx cyaneus Gyll, andSitona lineatus L. The latter appeared to be an efficient vector: acquisition and inoculation occurred at the first bite, the rate of transmission was c. 41%, and virus retention lasted for at least seven days.S. lineatus transmitted the virus from faba bean to lentil and pea, but not to the three genotypes of chickpea tested. This is the first report on the generaHypera, Pachytychius, andSmicronyx as virus vectors, and onA. radiolus, H. variabilis, P. strumarius, andS. cyaneus as vectors of BBMV.Out of 351 samples of food legumes with symptoms suggestive of virus infection, 16, 11, 19, and 17% of the samples of chickpea, lentil, pea, and common bean, respectively, were found infected when tested for BBMV in DAS-ELISA. This is the first report on the natural occurrence of BBMV in chickpea, lentil, pea, and common bean. The virus should be regarded as a food-legume virus rather than a faba-bean virus solely, and is considered an actual threat to food legume improvement programmes.     

Characterization of potato potyvirus Y (PVY) isolates from seed potato batches. Situation of the NTN, Wilga and Z isolates

A collection of 38 PVY isolates from seed potato batches, originating from several Western European countries, was characterized by using current biological, serological and molecular tools differentiating PVY strains and groups. The correlation between the three kinds of tests was good but not absolute. No single serological or PCR method was able to discriminate among the five isolate groups found. Twenty-nine isolates belonged to the PVY^N strain and six to the PVY^O strain. No PVY^C was found. Two other isolates reacted serologically like PVY^O, but were unable to elicit a hypersensitive response from the Ny_tbr gene and probably represent the PVY^Z group. At the molecular level, these two isolates showed a combination of both PVY^O and PVY^N and could be recombinants of these strains. Another isolate reacted serologically like PVY^O, but induced vein necrosis in tobacco, like PVY^N-Wilga. Some PVY^N isolates caused tuber ring necrosis in glasshouse conditions. These might belong to the PVY^NTN group. The PVY^NTN, PVY^N-Wilga and PVY^Z groups probably represent pathotypes within strains PVY^N and PVY^O, respectively. The present study also confirms previous reports showing a high genetic variation at the 5 end within the PVYN strain.     

The autumn leafroller: Phenology,damage and parasitoids in a Dutch apple orchard

The phenology of the autumn leafroller,Syndemis musculana, a local pest of apple, was studied in order to forecast larval emergence. From 1983–1986, peak flight as determined with sexpheromone traps was always between 13–18 May. The duration of embryonic development was determined at various constant temperatures and used to estimate the periods of egg hatch in these four years. Each year, most eggs should have hatched in the second decade of June.Differences in attack rates between apple cultivars seem to be explained largely by the variation in picking time. Larvae are only half grown at the beginning of harvest (cv. James Grieve), and have gone into hibernation when the latest variety (cv. Golden Delicious) is picked. Moreover, the varieties Cox's Orange Pippin and Belle de Boskoop, picked about half time, are liable to receive additional damage by caterpillars brought with the picked fruits into storage.Various hymenopterous parasites were reared from caterpillars. As the only leafroller in the orchard which hibernates as mature larva,S. musculana may promote winter survival of some parasitoids, like the eulophidColpyclypeus florus.Samenvatting De fenologie van de herfstbladroller (Syndemis musculana Hübner), een incidentele plaag op appel, werd nader bepaald met het doel het uitkomen van de eieren te kunnen voorspellen. In 1983–1986 viel de piekvlucht, bepaald met behulp van feromoonvallen, steeds tussen 13 en 18 mei.De ontwikkelingsduur van de eieren bij verschillende constante temperaturen werd gebruikt om de periode van uitkomen te schatten. De meeste eieren zullen ieder jaar in de eerste helft van juni uitkomen.Geconstateerde verschillen in schade tussen appelrassen blijken goeddeels terug te voeren op verschillen in pluktijdstip. De rupsen van de herfstbladroller zijn pas half-was als de eerste appels eind augustus geplukt worden, terwijl tegen het einde van de oogst begin oktober de meeste al in winterslaap zijn. Met name tussentijdse rassen, als Cox's Orange Pippin and Schone van Boskoop, lopen extra schade op doordat grotere rupsen met de geplukte vruchten in de kist terecht komen.Uit de rupsen werden negen, al van andere boomgaardbladrollers bekende, sluipwespen gekweekt, Omdat deze bladrollersoort, als enige in de boomgaard, als volgroeide rups overwintert, lijkt zij bij uitstek geschikt als winterwaard.This study was carried out at the Experimental Orchard De Schuilenburg, Schuilenburg 3, 4041 BK Kesteren, the Netherlands, to which address correspondence should be addressed.     

Characterization of Sclerotinia minor populations in Texas peanut fields

In the summer of 2004 an epidemic of sclerotinia blight of peanut, a disease caused by Sclerotinia minor, occurred in Texas in fields where the disease was never previously detected. The disease was observed on many plants within one of the fields (>3000 disease foci), although most foci were <1 m. It is hypothesized that these observations were inconsistent with the recent introduction of a monocyclic pathogen, even if disease developed under conducive environmental conditions. The pattern of disease is most suggestive of the presence of foliar (ascospore) infections, although air temperature was above the known limits for apothecia development if the pathogen had arrived in the field in 2004 peanut seed. To further examine this epidemic, 232 isolates were collected, across a variety of spatial scales spanning this field and other Texas peanut fields, and evaluated for aggressiveness, fungicide sensitivity and genotypic diversity. There was wide variation among isolates for the phenotypic characteristics measured, but there was no evidence that a genotypically unique, highly aggressive, and fungicide resistant isolate had been introduced or evolved. The predominant genotype, TX1, which contained 154 isolates, was found in every county and field population.     

Tumours of Begonia and some other ornamentals,induced by Corynebacterium fascians

In 1975 many tumours were observed in plants ofBegonia Schwabenland grown in Aalsmeer. Submersion of the roots ofNicotiana megalosiphon seedlings in a homogenate of tumorous tissue, induced tumours after two weeks. Short periods of submergence yielded results similar to those obtained after longer periods. Tumour homogenates lost their infectivity after ten min at 50°C. Aphids transmitted the infectious agent.Treatment with propylene oxide did not inhibit infectivity completely. Filtration through a 450 nm filter removed the infectious agent.Tobacco tumor virus or a viroid could not be isolated. Cultures ofCorynebacterium fascians, isolated from tumours ofN. megalosiphon were highly infectious and induced tumours in healthyN. megalosiphon andBegonia. Tumorous tissue homogenates ofPelargonium zonale, Dahlia sp.,Gladiolus sp., andLilium sp. also caused tumours inN. megalosiphon, from whichC. fascians was isolated. It was not possible to produce tumours inN. megalosiphon with homogenates from roses with symptoms of bud proliferation.Samenvatting In 1975 werden vele tumoren waargenomen inBegonia Schwabenland op Aalsmeerse bedrijven (Fig. 1). De infectiositeit van tumorweefsel kon goed en snel worden vastgesteld door de wortels van zaailingen vanNicotiana megalosiphon in een homogenaat van tumorweefsel te dompelen. Tumoren ontstonden na twee weken, de eindbeoordeling geschiedde na een maand (Fig. 2). Ook verschillende andereNicotiana spp.,Melilotus officinalis (Fig. 3) enPisum odoratum (Fig. 4) werden aangetast.Bij de infectiositeitstoets gaven zeer korte dompeltijden even goede resultaten als langere (Tabel 1). Infectieus sap verloor zijn infectievermogen na 10 min verhitting bij 50°C. Bladluizen brachten de smetstof over. Propyleenoxide verminderde de infectiositeit wel, doch onderdrukte deze niet totaal. Bij filtratie door een 450 nm filter bleef het infectieuse agens op het filter achter. Het tumor-inducerende agens was ook aanwezig in die delen van planten met tumoren welke gezond leken en het ging voor een gering deel over met zaad (Tabel 2).Uit tumoren konden wij geen tabakstumorvirus of een viroïde isoleren. Culturen vanCorynebacterium fascians, geïsoleerd uit tumoren vanN. megalosiphon bleken zeer infectieus en veroorzaakten tumoren inN. megalosiphon enBegonia. Homogenaten van tumorweefsel vanPelargonium zonale, dahlia (Fig. 5), gladiool (Fig. 6) enLilium Mid Century Hybrid Enchantment (Fig. 7) veroorzaakten ook tumoren opN. megalosiphon, waaruitC. fascians werd geïsoleerd. Met sap van kroeskopzieke rozen konden wijN. megalosiphon niet besmetten.     

Eyespot of Cereals Revisited: ITS phylogeny Reveals New Species Relationships

Four species so far classified in Pseudocercosporella or Ramulispora (hyphomycetes) are associated with eyespot disease symptoms of cereals. Two of these have been linked to teleomorphs that were described in Tapesia. Sequence data derived from the Internal Transcribed Spacer region (ITS1, 5.8S and ITS2) of the rDNA operon showed, however, that the eyespot fungi associated with Tapesia are not congeneric with Ramulispora sorghi, the type of Ramulispora. The genus name Tapesia is now rejected in favour of the conserved name Mollisia, which appears to comprise heterogeneous fungi. Tapesia yallundae is not closely related to the type of Mollisia, M. cinerea, but clusters separately, being more closely allied to species with Cadophora anamorphs. A new holomorph genus, Oculimacula, is therefore proposed for teleomorphs of the eyespot fungi, while the anamorphs are accommodated in Helgardia gen. nov.